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The thiol group of glutathione (GSH) was protected by 2, 4-dinitrochlorobenzene(DNCB). The product GSH-S-DNP was reacted with 2-methyl-1-propanol and 3-methyl-1-butanol respectively to obtain hapten6 and 7. Their structures were determined by 400 MHz1H NMR and IR. The two haptens were then covalently crosslinked with bovine serum albu-min(BSA) hy using glutaraldehyde as coupling agent to obtain antigen 6 and 7. The resultsof gel electrophoresis analysis gave the average molecular weight of 1. 27 ×10-22 kg for eachantigen. The average connected ratio of BSA to hapten was 1: 12. Other UV and fluores-cence spectral properties of the antigens are reported.